• Human vitamin D ELISA Kit
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Human vitamin D ELISA Kit

Human vitamin D ELISA Kit

Catalog Number: abs553813 Application: ELISA Reactivity: Human Conjugation:
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$450 USD
$450 USD
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Product Details

Product Specification

protein VD
Usage Specimen requirements:
1 . The specimen should be extracted as soon as possible after collection, and the extraction should be carried out according to the relevant literature.
Experiments should be carried out as soon as possible after extraction.
If the test cannot be carried out immediately, the specimen can be placed in -20℃ Store, but avoid repeated freezing and thawing
2 . Unable to detect containing NaN3 NaN3 Inhibition of horseradish peroxidase (HRP) activity.

Operation steps:
1. Dilution of standard: This kit provides one original standard, which can be diluted in a small test tube according to the following chart.
1600ug/L 5 150uL Addition of the original standard 150uL Standard dilution
800ug/L 4 No. Standard 150uL Of 5 150uL It is sufficient, for human peripheral blood, it is advisable to extend the lysis time to
400ug/L 3 150uL Of 4 No. Standard addition 150uL Standard dilution
200ug/L 2 No. Standard 150uL 3 No. Standard addition 150uL Standard dilution
100ug/L 1 No. Standard  150uL Of 2 No. Standard addition 150uL Standard dilution

2. Sample addition: Blank wells (no sample and enzyme labeled reagent are added to the blank control wells, and the other steps are the same), standard wells and sample wells to be tested are set respectively.
Accurate spike of standard on enzyme-labeled coated plate 50uL, first add sample diluent to the sample well to be tested 40uL, then add the sample to be tested 10uL (The final dilution of sample was 5 Times).
Add the sample to the bottom of the well of the enzyme labeled plate, try not to touch the well wall, and gently shake and mix well.
3. Incubation: After plate sealing with plate sealing film 37℃ Incubation 30 Minutes.
4. Solution preparation: will 30 Distilled water for double concentrated washing liquid 30 Time dilution for use
5. Washing: Carefully remove the sealing film, discard the liquid, spin dry, fill each well with washing liquid, and let stand 30 Discard it in seconds and repeat it 5 Times, pat dry.
6. Enzyme addition: enzyme labeled reagent is added to each well 50uL, except for blank holes.
7. Incubation: Same procedure 3.
8. Washing: Same operation 5.
9. Color development: Add color developer to each well first A 50uL And then adding a color developer B 50uL, gently shake and mix evenly, 37℃ Color development protected from light 10 Minutes.
10. Stop: Add stop solution per well 50uL, terminate the reaction (blue immediately turns yellow at this time).
11. Measurement: zeroing with blank holes, 450nm The absorbance of each well was measured sequentially by wavelength (OD Value).
The determination shall be performed after addition of the stop solution 15 Within minutes.

Calculation:
Taking the concentration of the standard as the abscissa, OD The value is the ordinate, draw a standard curve on the coordinate paper, and according to the sample OD The corresponding concentration is found out from the standard curve; Multiply by the dilution factor;
Or use the concentration of the standard substance to OD The linear regression equation of the standard curve is calculated from the value, and the sample OD The value is substituted into the equation, the sample concentration is calculated, and then multiplied by the dilution factor, which is the actual concentration of the sample.
Species Reactivity Human
Theory This kit uses double antibody sandwich method to determine the level of human vitamin D in specimens. Coating a microwell plate with purified human vitamin D antibody to make a solid phase antibody, adding vitamin D to the microwells coated with monoclonal antibody sequentially, and then combining with HRP-labeled vitamin D antibody to form an antibody-antigen-enzyme-labeled antibody complex, which is thoroughly washed and then added with substrate TMB to develop color. TMB is converted to blue under the catalysis of HRP enzyme and to the final yellow under the action of acid. There was a positive correlation between the depth of color and vitamin D in the sample. The absorbance (OD value) was measured with a microplate reader at a wavelength of 450nm, and the concentration of human vitamin D in the sample was calculated by the standard curve.
Source Human
Synonym Human VD ELISA Kit
Composition Kit composition:
Components Specifications
30 Double concentrated wash solution 20mL×1 Bottle
Enzyme labeled reagent 6mL×1 Bottle
Enzyme-labeled coated plate 12 Hole ×8 Strip
Sample dilution 6mL×1 Bottle
Color developer A Liquid 6mL×1 Bottle
Color developer B Liquid 6mL×1 Bottle
Stop liquid 6mL×1 Bottle
Standard ( 3200ug/L ） 0.5mL×1 Bottle
Standard dilution 1.5mL×1 Bottle
Sealing film 2 Zhang
Sealed bag 1 1
General Notes 1. The kit should be taken out of the refrigerated environment and balanced at room temperature for 1 hour before it can be used. If the enzyme-labeled coated plate is not used up after opening, the strips should be stored in a sealed bag.
2. Crystals may precipitate in the concentrated washing liquid. When diluting, it can be heated in a water bath to help dissolve, and the results will not be affected during washing.
3. A sampler should be used in each step of sampling, and its accuracy should be checked frequently to avoid test errors. It is best to control the sample addition time within 5 minutes. If the number of specimens is large, it is recommended to use a row gun to add samples.
4. Please make a standard curve at the same time of each measurement, preferably a double hole. If the content of the substance to be tested in the sample is too high (the OD value of the sample is greater than the OD value of the first well of the standard well), please dilute it with the sample diluent for a certain multiple (n times) before measuring, and please finally multiply it by the total dilution multiple (× n × 5) when calculating.
5. The sealing film is only for one-time use to avoid cross-contamination.
6. Please keep the substrate away from light.
7. Strictly follow the instructions, and the test results must be determined based on the reading of the microplate reader.
8. All samples, washing solutions and various wastes should be treated as infectious agents.
9. Components of different batch numbers of this reagent shall not be mixed.
Storage Temp. Store at 2-8 °C, shelf life 6 months.
Test Range 50ug/L -1600ug/L
Applications It is used to determine the content of vitamin D in human serum, plasma and related fluid samples.